• Protocol for engineering bindi...

Treffer: Protocol for engineering binding domains to recognize ligand-bound receptors by using yeast surface display.

Title:
Protocol for engineering binding domains to recognize ligand-bound receptors by using yeast surface display.
Authors:
Dobersberger M; Department of Chemistry, Institute of Biochemistry, BOKU University, Vienna 1190, Austria., Sumesgutner D; Department of Chemistry, Institute of Biochemistry, BOKU University, Vienna 1190, Austria; CD Laboratory for Next Generation CAR T Cells, Vienna 1090, Austria. Electronic address: delia.sumesgutner@boku.ac.at., Zajc CU; Department of Chemistry, Institute of Biochemistry, BOKU University, Vienna 1190, Austria; CD Laboratory for Next Generation CAR T Cells, Vienna 1090, Austria., Traxlmayr MW; Department of Chemistry, Institute of Biochemistry, BOKU University, Vienna 1190, Austria; CD Laboratory for Next Generation CAR T Cells, Vienna 1090, Austria. Electronic address: michael.traxlmayr@boku.ac.at.
Source:
STAR protocols [STAR Protoc] 2024 Dec 20; Vol. 5 (4), pp. 103339. Date of Electronic Publication: 2024 Sep 24.
Publication Type:
Journal Article; Research Support, Non-U.S. Gov't
Language:
English
Journal Info:
Publisher: Cell Press Country of Publication: United States NLM ID: 101769501 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 2666-1667 (Electronic) Linking ISSN: 26661667 NLM ISO Abbreviation: STAR Protoc Subsets: MEDLINE
Imprint Name(s):
Original Publication: [Cambridge, MA] : Cell Press, [2020]-
MeSH Terms:
Protein Engineering*/methods , Saccharomyces cerevisiae*/genetics , Saccharomyces cerevisiae*/metabolism, Ligands ; Flow Cytometry/methods ; ErbB Receptors/metabolism ; ErbB Receptors/genetics ; Epidermal Growth Factor/metabolism ; Cell Surface Display Techniques/methods ; Protein Binding ; Peptide Library ; Humans
Contributed Indexing:
Keywords: biotechnology and bioengineering; cancer; flow cytometry
Substance Nomenclature:
0 (Ligands)
EC 2.7.10.1 (ErbB Receptors)
62229-50-9 (Epidermal Growth Factor)
0 (Peptide Library)
Entry Date(s):
Date Created: 20240925 Date Completed: 20241224 Latest Revision: 20250610
Update Code:
20250611
PubMed Central ID:
PMC11462054
DOI:
10.1016/j.xpro.2024.103339
PMID:
39321026
Database:
MEDLINE

Weitere Informationen

Yeast surface display is a versatile protein engineering technology, enabling precise control of the applied selection pressure. We present a yeast-surface-display-based protocol for the enrichment of binders specifically recognizing ligand-bound receptors. We describe steps for magnetic bead selections, random mutagenesis, and flow cytometric sorting, followed by library sequencing and detailed analysis of enriched clones. While this approach is exemplified with rcSso7d-based libraries and epidermal growth factor (EGF)-epidermal growth factor receptor (EGFR) complexes, it can also be adapted to other binder scaffolds and ligand-receptor systems. For complete details on the use and execution of this protocol, please refer to Dobersberger et al. <sup>1</sup> .
(Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Declaration of interests M.W.T. receives funding from Miltenyi Biotec.